Fish collagen extraction

and application
Nguyen Hoang Dung, Phan Dinh Tuan, Mai Thanh Phong,
Nguyen Thi Nguyen, Ngo Hong Bao Chau, Nguyen Thi
Diem Phuong, Doan Huu Luc

Collagen
What is collagen?
Collagen: the main protein of connective tissue in
animals and the most abundant protein in mammals,
making up about 25% to 35% of the whole-body protein
content. Collagen constitutes 1% to 2% of muscle tissue,
and accounts for 6% of the weight of strong, tendinous
muscles.
What is it used for?
Food: structure additive,
Pharmaceutical: soft and hard capsule, artificial skin,
Cosmetic: repair rough skin, wrinkle-resistant,

Main sources
Pig skins or cow skins and bones: low cost of the final

product

Why do we shift to collagen extraction from fish?

Major advantage of marine collagen sources is that they are not

associated with the risk of bovine spongiform encephalopathy

(BSE) outbreaks.
Fish collagen is acceptable for Islam and with a minimum
restriction for Judaism.
Fish skin is a major by-product of the fish-processing industry,
causing wastage and pollution, and could provide a valuable source
of collagen.
More suitable for human skin than collagen from livestock and
poultry.

Extraction methods
Chemical methods:

Solvents for removing non-collagenous proteins, defatting, deordoring: salt, dilute acid, alcohol,
detergents, H2O2
Chemicals for deashing: EDTA ( for pre-treating bone)
Solvent for extracting collagen: delute acide (acid acetic, acide lactic), enzyme (pepsin), bacteria
Bacillus, yeast. Temperature: 0-40C

Our researching
1.Pre-treatment steps:
2. Extracting acid-soluble collagen
3.Purify collagen

Source: Min Zhang,Wentao Liu, Guoying Li,(2008),Sichuan University, China

Source: Inwoo Bae,(2007),Nagasaki University, Japan
Source: Takeshi Nagai,(1999),Kyushu University, Japan

Extraction methods
Supercritical extraction method by using CO2 as

solvent

Environmental improvement and reduced product contamination

Advantages
Easy to control dissolving power of supercritical fluid

(SCF) by controlling T, P.
Easy to recover SCF by decompressing pressure.
Impossible to separate the precipitate from extracts by
centrifugation.
Non- toxic solvent
Applicable to extract the thermally decomposed
compounds

Disadvantages
Require high pressure.
Investing elaborate equipment and high capital.
High technical worker.

ANALYSE METHODS
1. Sodium dodecyl sulphate polyacrylamine gel

electrophoresis (SDS -PAGE) to derterminate the

forms of collagen (-chain,-chain)
2. SDS- PAGE to dertermine Peptide mapping
3. Determination of hydroxyproline content
4. Determination of denaturation temperature
5. Viscosity of collagen solution
6. Effect pH and NaCl on Collagen solubility
7. FT-IR
8.SEM to determine structure of collagen

Source: Boyl R. Switzer,(1991),University of North Carolina, USA.

Methods to be applied
Data-base set up, investigation of structure,

characteristics and application capability of collagen

Investigation of collagen concentration in skin and
skeleton of catfish (basa fish).
Choice of suitable solvent for collagen extraction
Implementation of extraction process for high yield
and selection of collagen extraction.
Analysis and evaluation of extracted collagen:

RBE Plan
Supervising system of masters program
Project team:
Dr. Nguyen Hoang Dzung
Dr. Mai Thanh Phong
Prof. Dr. Phan Dinh Tuan
Dr. Doan Huu Luc
KS. Nguyen Thi Nguyen

Le Thi Thu Huong,

PhD. student

Ngo Hong Bao Chau

Master student

Nguyen Thi
Diem Phuong
Undergraduate student

RBE Plan
For the leader of the laboratory:
Supervising master students from the first day of the
program until they finish it.
Guiding/supervising master students so that they can
create one academic paper based on the joint research
under SUPREM-HCMUT.
Giving regular lab meetings where master students are
required to report their research progress.

RBE Plan
For master students:
Must create their master thesis based on the research
outcomes.
Requested to write an academic paper as the first author
and based on the Key Issues given.
Substantial goals:
One PhD, one master student and 2 undergraduate
students involve in the research work along with lab
management and team leader.
One master thesis will be created on July 2010.
At least two papers conducted by master student will be
submitted.

RBE Plan
Actions to achieve the substantial goal:
Assigning a supervisor to the master student
Organizing regular seminars within the research group
to transform the content of some lectures to research
based seminars
Desk arrangement in the Lab for students
Assigning a research assistant to master students
Planning the research project taking the publication
into consideration
Frequent research discussions will be once a week

References
Phanat Kittiphattanabawon et al,(2004). Characterisation of acide-

soluble collagen from skin and bone of bigeye snapper, Prince of

Songkla University, Thailand.
Boyl R. Switzer,(1991),Dertermination of hydroxyproline, University of
North Carolina, USA.
Min Zhang,Wentao Liu, Guoying Li,(2008), Isolation and
characterisation of collagen from the skin of largefin longbarbel catfish,
Sichuan University, Chin.
Inwoo Bae,(2007), Biochemical properties of acid-soluble collagens
extracted from the skin of underutilised fishes, Nagasaki University,
Japan
Takeshi Nagai et al,(1999), Isolation of collagen from fish waste
material-skin, bone, and fins, Kyushu University, Japan.
Losso et al, Extraction of collagen from calcified tissues, Patent No.
US2005/0267292 A1, United States.
Hsiao et al, Novel collagen extraction method, Patent No.
US2004/0253678 A1, United States.